title: Quantitative determination of 5-aminolaevulinic acid and its esters in cell lysates by HPLC-fluorescence.
abstract: The development of a reliable sensitive method for the HPLC determination of 5-aminolaevulinic acid (ALA) and ALA esters in cell lysates is described. The method relies on the quantification of a fluorescent derivative of ALA following its derivatisation with acetylacetone and formaldehyde. Following this procedure it is possible to quantify ALA in cell lysates with no need for pre-purification of the sample. The method has been validated in two ranges of concentration (0.6-65 microM, 0.1-10 microg/mL, and 30-600 microM, 5-100 microg/mL), and has also been extended and validated for the determination of ALA released from ALA prodrugs after acidic hydrolysis.
authors.affiliation: Wolfson Laboratory of Medicinal Chemistry, Department of Pharmacy and Pharmacology, University of Bath, Bath, UK.
authors.forename: Francesca
authors.initials: F
authors.lastname: Giuntini
authors.forename: Ludovic
authors.initials: L
authors.lastname: Bourré
authors.forename: Alexander J
authors.initials: AJ
authors.lastname: Macrobert
authors.forename: Michael
authors.initials: M
authors.lastname: Wilson
authors.forename: Ian M
authors.initials: IM
authors.lastname: Eggleston
mesh_terms: D000622:Aminolevulinic Acid / Q000032:analysis* / Q000378:metabolism; D000704:Analysis of Variance; D000818:Animals; D002460:Cell Line; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D004951:Esterification; D004952:Esters / Q000032:analysis; D015603:Keratinocytes / Q000737:chemistry* / Q000378:metabolism; D016014:Linear Models; D051379:Mice; D011355:Prodrugs / Q000378:metabolism; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity
chemical_list: D004952:Esters; D011355:Prodrugs; D000622:Aminolevulinic Acid